pET-59-DEST

Gateway® Nova bacterial destination vector for expressing proteins tagged at the N-terminus with thioredoxin and at the C-terminus with Strep-Tag II.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidsGateway® Cloning VectorspET-59-DEST
AvrII (7097) BstBI (7078) Strep-Tag II SalI (6924) PstI (6922) BfuAI - BspMI (6911) BmgBI (6701) SmaI - SrfI (6667) TspMI - XmaI (6665) BbvCI (6521) PasI (6028) EcoRI (5796) EagI - NotI (5476) PmlI (5338) thrombin site SacI (5328) Eco53kI (5326) RsrII (5095) BmtI (5000) NheI (4996) NdeI (4991) 6xHis ATG NcoI (4955) RBS XbaI (4916) T7 promoter SgrAI (4805) SphI (4657) EcoNI (4592) BstAPI (4448) BclI * (4110) BstEII (3942) ApaI (3921) PspOMI (3917) BlpI (7115) DraIII (245) PsiI (370) AhdI (655) BsaI (716) BglI (775) FspI (877) PvuI (1025) PciI (2260) BspQI - SapI (2377) PflFI - Tth111I (2518) PpuMI (3255) HpaI (3622) pET-59-DEST™ 7198 bp
AvrII  (7097)
1 site		 C C T A G G G G A T C C
BstBI  (7078)
1 site		 T T C G A A A A G C T T
SalI  (6924)
1 site		 G T C G A C C A G C T G
PstI  (6922)
1 site		 C T G C A G G A C G T C
BfuAI  (6911)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (6911)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
BmgBI  (6701)
1 site		 C A C G T C G T G C A G
This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
SmaI  (6667)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
SrfI  (6667)
1 site		 G C C C G G G C C G G G C C C G
TspMI  (6665)
1 site		 C C C G G G G G G C C C
XmaI  (6665)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
BbvCI  (6521)
1 site		 C C T C A G C G G A G T C G
PasI  (6028)
1 site		 C C C W G G G G G G W C C C
Sticky ends from different PasI sites may not be compatible.
EcoRI  (5796)
1 site		 G A A T T C C T T A A G
EagI  (5476)
1 site		 C G G C C G G C C G G C
NotI  (5476)
1 site		 G C G G C C G C C G C C G G C G
PmlI  (5338)
1 site		 C A C G T G G T G C A C
SacI  (5328)
1 site		 G A G C T C C T C G A G
Eco53kI  (5326)
1 site		 G A G C T C C T C G A G
RsrII  (5095)
1 site		 C G G W C C G G C C W G G C
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BmtI  (5000)
1 site		 G C T A G C C G A T C G
NheI  (4996)
1 site		 G C T A G C C G A T C G
NdeI  (4991)
1 site		 C A T A T G G T A T A C
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
NcoI  (4955)
1 site		 C C A T G G G G T A C C
XbaI  (4916)
1 site		 T C T A G A A G A T C T
SgrAI  (4805)
1 site		 C R C C G G Y G G Y G G C C R C
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
SphI  (4657)
1 site		 G C A T G C C G T A C G
EcoNI  (4592)
1 site		 C C T N N N N N A G G G G A N N N N N T C C
The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BstAPI  (4448)
1 site		 G C A N N N N N T G C C G T N N N N N A C G
Sticky ends from different BstAPI sites may not be compatible.
BclI  (4110)
1 site		 T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BstEII  (3942)
1 site		 G G T N A C C C C A N T G G
Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
ApaI  (3921)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
PspOMI  (3917)
1 site		 G G G C C C C C C G G G
BlpI  (7115)
1 site		 G C T N A G C C G A N T C G
Sticky ends from different BlpI sites may not be compatible.
DraIII  (245)
1 site		 C A C N N N G T G G T G N N N C A C
Sticky ends from different DraIII sites may not be compatible.
PsiI  (370)
1 site		 T T A T A A A A T A T T
AhdI  (655)
1 site		 G A C N N N N N G T C C T G N N N N N C A G
The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BsaI  (716)
1 site		 G G T C T C N C C A G A G N ( N ) 4
Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BglI  (775)
1 site		 G C C N N N N N G G C C G G N N N N N C C G
Sticky ends from different BglI sites may not be compatible.
FspI  (877)
1 site		 T G C G C A A C G C G T
PvuI  (1025)
1 site		 C G A T C G G C T A G C
PciI  (2260)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (2377)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different BspQI sites may not be compatible.
SapI  (2377)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
PflFI  (2518)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (2518)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
PpuMI  (3255)
1 site		 R G G W C C Y Y C C W G G R
Sticky ends from different PpuMI sites may not be compatible.
HpaI  (3622)
1 site		 G T T A A C C A A T T G
lacI
3397 .. 4479  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lacI
3397 .. 4479  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
AmpR
582 .. 1442  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   582 .. 1373  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
582 .. 1442  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1374 .. 1442  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
582 .. 1442  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
5583 .. 6242  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
5583 .. 6242  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
1616 .. 2204  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1616 .. 2204  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
12 .. 467  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
12 .. 467  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
ATG
4957 .. 4959  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
4957 .. 4959  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
4969 .. 4986  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
4969 .. 4986  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
TrxA
4993 .. 5322  =  330 bp
110 amino acids  =  11.9 kDa
Product: E. coli thioredoxin
TrxA
4993 .. 5322  =  330 bp
110 amino acids  =  11.9 kDa
Product: E. coli thioredoxin
thrombin site
5329 .. 5346  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
thrombin site
5329 .. 5346  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
ccdB
6584 .. 6889  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.
ccdB
6584 .. 6889  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.
rop
2634 .. 2825  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
2634 .. 2825  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
attR1
5350 .. 5474  =  125 bp
recombination site for the Gateway® LR reaction
attR1
5350 .. 5474  =  125 bp
recombination site for the Gateway® LR reaction
attR2
6930 .. 7054  =  125 bp
recombination site for the Gateway® LR reaction
attR2
6930 .. 7054  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
1443 .. 1535  =  93 bp
AmpR promoter
1443 .. 1535  =  93 bp
lacI promoter
4480 .. 4557  =  78 bp
lacI promoter
4480 .. 4557  =  78 bp
T7 terminator
7126 .. 7173  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
7126 .. 7173  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac UV5 promoter
5499 .. 5529  =  31 bp
3 segments
   Segment 1:  -35  
   5499 .. 5504  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
5499 .. 5529  =  31 bp
3 segments
   Segment 2:  
   5505 .. 5522  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
5499 .. 5529  =  31 bp
3 segments
   Segment 3:  -10  
   5523 .. 5529  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
5499 .. 5529  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
lac operator
4889 .. 4913  =  25 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
4889 .. 4913  =  25 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
Strep-Tag II
7062 .. 7085  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an engineered form of streptavidin
Strep-Tag II
7062 .. 7085  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an engineered form of streptavidin
T7 promoter
4870 .. 4888  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4870 .. 4888  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
4944 .. 4949  =  6 bp
ribosome binding site
RBS
4944 .. 4949  =  6 bp
ribosome binding site
ORF:  712 .. 978  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2857 .. 3225  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  4957 .. 5385  =  429 bp
ORF:  142 amino acids  =  15.5 kDa
ORF:  4655 .. 4906  =  252 bp
ORF:  83 amino acids  =  8.6 kDa
ORF:  6584 .. 6889  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  3363 .. 3614  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  4518 .. 4781  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  5583 .. 6242  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  3377 .. 3640  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  4229 .. 4729  =  501 bp
ORF:  166 amino acids  =  17.5 kDa
ORF:  3397 .. 4356  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  582 .. 1442  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2634 .. 2858  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
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Download pET-59-DEST.dna file

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Individual Sequences & Maps

pAd BLOCK-iT-DEST
pDEST R4-R3 Vector II
pENTR11
pHELLSGATE 4
pAd CMV V5-DEST
pDEST10
pENTR1A
pHELLSGATE 8
pAd PL-DEST
pDEST14
pENTR2B
pIB V5-His-DEST
pBAD-DEST49
pDEST15
pENTR3C
pLenti4 BLOCK-iT-DEST
pcDNA-DEST40
pDEST17
pENTR4
pLenti4 V5-DEST
pcDNA-DEST47
pDEST20
pET-53-DEST
pLenti6.2 C-Lumio V5-DEST
pcDNA-DEST53
pDEST24
pET-54-DEST
pLenti6.2 N-Lumio V5-DEST
pcDNA3.1 nV5-DEST
pDEST26
pET-55-DEST
pLenti6.2 V5-DEST
pcDNA3.2 capTEV-CT V5-DEST
pDEST27
pET-56-DEST
pLenti6.3 TO V5-DEST
pcDNA3.2 capTEV-NT V5-DEST
pDEST8
pET-57-DEST
pLenti6.3 V5-DEST
pcDNA3.2 V5-DEST
pDONR221 P5-P2
pET-58-DEST
pLenti6.4 R4R2 V5-DEST
pcDNA6.2 C-EmGFP-DEST
pDONR P2r-P3
pET-59-DEST
pLenti6 BLOCK-iT-DEST
pcDNA6.2 C-YFP-DEST
pDONR P4-P1r
pET-60-DEST
pLenti6 capTEV-CT-DEST
pcDNA6.2 cGeneBLAzer-DEST
pDONR Zeo
pET-61-DEST
pLenti6 capTEV-NT-DEST1
pcDNA6.2 cLumio-DEST
pDONR201
pET-62-DEST
pLenti6 UbC V5-DEST
pcDNA6.2 EmGFP-Bsd V5-DEST
pDONR207
pET-DEST42
pLenti6 V5-DEST
pcDNA6.2 N-EmGFP-DEST
pDONR221
pET104-DEST
pLenti7.3 V5-DEST
pcDNA6.2 N-YFP-DEST
pDONR221 P1-P4
pET104.1-DEST
pMT-DEST48
pcDNA6.2 nGeneBLAzer-DEST
pDONR221 P1-P5r
pET160-DEST
pMT BioEase-DEST
pcDNA6.2 nLumio-DEST
pDONR221 P3-P2
pET161-DEST
pSCREEN-iT lacZ-DEST
pcDNA6.2 V5-DEST
pDONR221 P4r-P3r
pEXP1-DEST
pT-REx-DEST30
pcDNA6.2 V5-pL-DEST
pDONR221 P5-P4
pEXP2-DEST
pT-REx-DEST31
pCMV SPORT6
pDONR222
pEXP3-DEST
pYES-DEST52
pCMV SPORT6.1
pDONR223
pEXP4-DEST
Reading Frame Cassette A
pCOLA-2-DEST
pEF-DEST51
pHELLSGATE
Reading Frame Cassette B
pCOLA-3-DEST
pEF5 FRT V5-DEST
pHELLSGATE 12
Reading Frame Cassette C.1
pDEST R4-R3
pENTR GeneBLAzer

Plasmid Sets

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Plant Vectors
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Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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